观赏番茄观赏茄子双向嫁接研究

利用观赏茄子的一个分枝作砧木,观赏番茄作接穗;观赏番茄的一个侧枝作砧木,观赏茄子作接穗进行观赏番茄观赏茄子双向嫁接研究。试验结果表明,观赏番茄作接穗时嫁接成活率明显高于观赏茄子作接穗时的嫁接成活率,能否嫁接成活与嫁接时的砧木、接穗的茎粗有一定的关系,本试验中观赏番茄、观赏茄子茎粗在0.5 cm时嫁接成活率最高。     

两种鹅膏菌纯培养菌丝的分离培养和生长形态

采集并组织分离到假褐云斑鹅膏(Amanita pseudoporphyria)和块鳞鹅膏(Amanita spissa)的纯培养菌丝.对2株鹅膏菌的培养发现虽然分离成功率很低,但是菌丝培养相对容易,在普通琼脂培养基上生长良好;2种鹅膏菌菌丝的生长形态和方式存在较大差别,对氨基酸的利用也存在差别,但是气生菌丝的延伸速率却非常相似.液体培养条件下,假褐云斑鹅膏菌丝体干重达到0.726mg·mL-1,块鳞鹅膏干重达到0.424mg·mL-1.     

Irbesartan alleviates hepatic steatosis in db/db mice by inducing auto-phagy

AIM: To investigate the effect of irbesartan on the fatty liver of db/db mice and whether autophagy is involved in the process. METHODS: Male db/db mice (n=24) were randomly divided into model group and irbesartan group, and 12 db/m mice with similar age and weight were selected as normal control group. After 16 weeks of intervention respectively, the fatty liver-related parameters including body weight, liver index, blood lipid, liver function and pathological changes in the liver were observed. The protein levels of p-PI3K, p-Akt, and p-mTOR, as well as Atg-7, beclin-1 and LC3B in the liver tissues were detected by Western blot, and the autophagosomes in the liver were observed under electron microscope. RESULTS: Compared with the model group, the body weight, liver index, blood lipids, alanine and aspartate aminotransferase were decreased in irbesartan group (P<0.05). Moreover, the pathological changes in the liver were significantly ameliorated in irbesartan group than that of model group. Importantly, the protein levels of p-PI3K, p-Akt and p-mTOR were decreased with irbesartan administration, while the expression of Atg-7, beclin-1 and LC3B-Ⅱ was increased(P<0.05), which resulted in a distinct increase in autophagosomes. CONCLUSION: Irbesartan alleviates hepatic steatosis in db/db mice by inhibiting the PI3K/Akt/mTOR signaling pathway and upregulating the protein expression of Atg-7, beclin-1 and LC3B-Ⅱ, thereby inducing autophagy in hepatocytes.     

Adeno-associated virus type 9-mediated p65 silencing inhibits angiotensin II-induced apoptosis of H9c2 cells

AIM: To study the effect of p65 gene silencing by adeno-associated virus type 9 (AAV9)-mediated RNA interference on angiotensin Ⅱ (Ang Ⅱ; 10^-6 mol/L for 24 h)-induced apoptosis of rat ventricular H9c2 myocytes, and to elucidate the possible mechanism. METHODS: The H9c2 cells were transfected with rAAV9-eGFP and rAAV9-eGFP-NF-κB p65-siRNA at multiplicity of infection (MOI)=4×10⁶ vg/cell. eGFP expression in the cells was observed under an inverted fluorescence microscope, and the percentage of eGFP positive cells was determined by flow cytometry. The expression of p65 was determined by Western blot. CCK-8 assay was used to measured the viability of transfected H9c2 cells. The apoptosis of the cells transfected with the virus and with Ang Ⅱ stimulation was analyzed by flow cytometry. RESULTS: The cells began to exhibit eGFP expression on the 2nd day after transfection. The fluorescence intensity was increased over the time of transfection. eGFP expression reached the maximum on the 5th day, and the transfection efficiency was (52.7±1.9)% at this time point. Compared with blank control group, no significant effect of AAV9 on the viability of H9c2 cells was observed. In resting state, p65 in the H9c2 cells had a certain activity. After Ang Ⅱ stimulation, the activity of p65 was obviously increased, while transfection of rAAV9-eGFP-NF-κB p65-siRNA effectively inhibited the expression of p65. The apoptosis of H9c2 cells in Ang Ⅱ stimulation group was significantly higher than that in blank control group, while transfection of rAAV9-eGFP-NF-κB p65-siRNA effectively inhibited apoptosis of H9c2 cells. CONCLUSION: Transfection of rAAV9-eGFP-NF-κB p65-siRNA effectively inhibits the expression of p65 gene of NF-κB pathway in the H9c2 cells without causing cell growth inhibition, and reduces the apoptosis induced by Ang Ⅱ.     

京冀蔬菜均衡供应分析及对策建议

北京蔬菜自给率偏低,随着城乡居民对"菜篮子"产品的需求日益提高,加强"菜篮子"区域间合作的需求已日益迫切。而京冀蔬菜产业联系紧密,在京津冀协同发展背景下,开展蔬菜生产与流通调研,加强两地供需对接,具有重要现实意义。该研究在对河北省环京蔬菜生产大县调研基础上,从供求数量、结构、质量、渠道4个方面对京冀蔬菜均衡供应进行对比分析,并针对京冀蔬菜均衡供应在体制机制、生产、流通、销售等环节存在的问题,从政策机制、高端生产、流通网络、信息服务方面提出保障京冀蔬菜均衡供应的有关对策建议。以期达到推动京冀区域农产品产销对接和"菜篮子"供应保障的目的。     

阳台水培苦苣营养液配方筛选

在阳台上水培苦苣,通过测量苦苣的形态指标、叶绿素、产量和品质等指标,以期筛选出比较适合阳台水培苦苣的营养液配方。结果表明,经郑州市蔬菜研究所的营养液配方处理的阳台水培苦苣株高、叶片数、叶面积、叶绿素含量、产量、VC含量、可溶性糖含量、总蛋白质含量均显著高于其他各处理,硝酸盐含量较低。     

蓝狐MITF-M基因序列扩增及生物信息学分析

试验旨在研究蓝狐MITF-M基因核心启动子活性区、转录因子结合位点、基因编码蛋白结构及功能,为探究该基因的表达调控机制提供理论依据。从成年蓝狐背部皮肤组织中采集1 cm×1 cm样品,采用RT-PCR扩增及测序技术首次获得MITF-M基因序列3 880 bp（包括5'侧翼区2 262 bp、CDS区1 260 bp、3'侧翼区358 bp）,编码419个氨基酸残基。生物信息学分析结果显示,蓝狐MITF-M基因5'侧翼区存在潜在的启动子区域（-86~-336 bp）,且发现TATA框、CAAT框、GC框和CRE等调控元件及CREB、LSF和Sp1等多种转录因子。MITF-M基因编码的蛋白是定位于细胞核和细胞质的不稳定、可溶性亲水蛋白质。亚细胞定位结果提示该蛋白在细胞核、细胞质和线粒体的概率较高,分别为65.2%、17.4%和8.7%。DNAMAN软件对MITF-M基因编码蛋白的二级结构预测发现,该蛋白由α螺旋（33.66%）、β折叠（16.66%）和无规卷曲（49.68%）组成,且不存在跨膜区域和信号肽。蓝狐与其他物种MITF-M基因编码的氨基酸序列同源性对比和系统进化树分析均提示蓝狐与犬的遗传亲缘关系最近,与哺乳动物和禽类均具有较高的同源性（>89.1%）,说明MITF-M基因编码区在物种进化过程中较为保守。本研究为深入研究MITF-M基因调控狐狸被毛颜色的分子遗传机制奠定了理论基础。     

Role of PI3K/Nrf2 signaling pathway in endotoxin-induced acute kidney injury in rabbits

AIM: To evaluate the role of phosphatidylinositol 3-kinase/nuclear factor E2-related factor 2 (PI3K/Nrf2) signaling pathway in endotoxin-induced acute kidney injury in rabbits. METHODS: Healthy male New Zealand white rabbits were randomly divided into 5 groups: control group (group C), LPS group (group L), wortmannin+LPS group (group WL), wortmannin group (group W) and dimthyl sulfoxide (DMSO) group (group D). Wortmannin at dose of 0.6 mg/kg was injected via the auricular vein in groups W and WL, DMSO at concentration of 0.08 mL/kg was injected in group D, while normal saline (0.08 mL/kg) was injected in groups C and L. LPS at dose of 5 mg/kg was injected via the auricular vein in groups L and WL 30 min later, and equal volume of normal saline was injected in group C, D and W for control. The rabbits were sacrificed 6 h after LPS or normal saline administration. The kidneys were removed for microscopic examination and the determination of histological scores of kidney (HSK). The concentrations of blood urea nitrogen (BUN) and creatinine (Cr), urinary α1-microglobulin (α1-MG), MDA content, SOD activity, the mRNA expression of Nrf2 and HO-1, and the protein levels of total Akt, p-Akt, total Nrf2, p-Nrf2, nuclear Nrf2 and HO-1 in the renal tissues were also detected. RESULTS: Compared with groups C, D and W, the concentrations of BUN and Cr, urinary α1-MG concentration, MDA content and HSK were significantly increased, while SOD activity was significantly decreased (P<0.05). The mRNA expression of Nrf2 and HO-1, and the protein levels of p-Akt, total Nrf2, p-Nrf2, nuclear Nrf2 and HO-1 in the renal tissues were significantly increased in groups L and WL. No significant change among groups C, D and W was observed. Compared with group L, the concentrations of BUN and Cr, urinary α1-MG concentration, MDA content and HSK were significantly increased, while SOD activity, the mRNA expression of Nrf2 and HO-1, and the protein levels of p-Akt, total Nrf2, p-Nrf2, nuclear Nrf2 and HO-1 in the renal tissues were significantly decreased in group WL. CONCLUSION: Activation of PI3K/Nrf2 signaling pathway may be one of the regulatory mechanisms of the body adapting to the endotoxin-induced acute kidney injury in rabbits.     

Microarray-based miRNAs profiling in lower limb arteriosclerosis of diabetic rats

AIM: To establish the profiling of microRNAs (miRNAs) in the lower extremity arterial tissue between diabetic rats with lower limb arteriosclerosis (DAS) and diabetic rats with normal lower limb (DN), and to explore the possible molecular mechanisms involved in aberrant miRNA expression in DAS. METHODS: The rat models of DAS and DN were successfully established. The respective lower extremity arterial tissue was isolated. The total miRNAs were purified for a hybridization detection by miRNA microarray. The results of chip scanning and data were analyzed and verified by RT-qPCR. RESULTS: Ten miRNAs related to DAS, including rno-miR-206-3p, rno-miR-133a-5p, rno-miR-133b-3p, rno-miR-133a-3p, rno-miR-325-5p, rno-miR-675-3p, rno-miR-411-5p, rno-miR-329-3p, rno-miR-335 and rno-miR-126a-3p, were determined. All 10 abnormally expressed miRNAs were up-regulated. The validating results of RT-qPCR confirmed 9 of the miRNAs in line with chip expression. Just rno-miR-335 showed the opposite between PCR detection and microarray result. CONCLUSION: A group of miRNAs in diabetic rats suffering from lower limb arteriosclerosis plays an important role in the vascular atherosclerosis process. The abnormal expression of miRNAs is likely to affect the vascular atherosclerosis process.     

piggyBac转座子介导的家蚕细胞转基因研究初探

为探讨稳定转化家蚕细胞表达外源基因的新方法,以家蚕核型多角体病毒(BmNPV)极早期蛋白基因(ie-1)启动子元件驱动新霉素抗性基因(neor),并克隆至具有绿色荧光蛋白基因(gfp)标记的昆虫转座子载体piggyBac中,构建转基因载体pigA3-IE-Neo。以该载体转染家蚕BmN细胞,用终浓度800μg/mL的遗传霉素(geneticin,G418)筛选3个月,获得了稳定转化的细胞,呈现绿色荧光的细胞数达80%以上。通过PCR鉴定证实了细胞基因组DNA中neor基因和gfp基因的存在。